| Objective The study was aimed to establish a rat model of breast tumor induced by subcutaneous injection of 7,12-dimethylbenz(a)anthracene,and to give the aplysin intervention.The inhibitory effect of aplysin on rat breast tumor was evaluated by the incidence of tumor,tumor inhibition rate in each group.To investigate the protective effect of aplysin on intestinal barrier injury induced by DMBA in rats with breast tumor.Methods1.Establishment of animal model:After one week adaptation with general meals,60 Female Spraque-Dawley rats were randomly divided into normal control group,model group and aplysin intervention group.In addition to the normal control group,the other two groups were injected subcutaneously with the same amount of Tween-80 into the right buttocks of rats.The normal control group was injected with the same amount of Tween-80.From the next day,the aplysin intervention group was given 40 mg/kg intragastrically,and the normal control group and model group were given the same amount of soybean oil intragastrically.2.Pathological diagnosis of tumor tissue: the tumor tissue of rats in model group and aplysin intervention group was fixed with 10% formaldehyde,embedded in paraffin,sectioned,dehydrated,stained with HE,and the morphological changes of each group were observed by optical microscope.3.The incidence of breast cancer,tumor latency,average tumor weight,tumor inhibition rate and organ index were calculated.4.HE staining was used to evaluate the pathology of jejunum in rats.5.The concentrations of IL-4,IL-6,IL-10 and TNF-α in rat plasma were detected6.The levels of D-LA and DAO in rat plasma were detected by ELISA.7.Western blot was used to observe the expression of tight junction protein occludin and ZO-1 in rat jejunum.8.Real-Time PCR was used to quantitatively analyze the variable region of 16 s RDNA V3 in rat fecal intestinal flora.Results1.No tumor was found in the normal control group.The average latency of tumor was shorter in the model group,significantly prolonged in the aplysin group,the tumor inhibition rate was 49.83%,and the thymus index was significantly increased.2.HE staining showed that compared with the normal control group,the intestinal wall of the model group was damaged,the number of intestinal villi decreased,the length of intestinal villi became shorter,most of the villi expanded and fused,and the top appeared defect.The morphology of intestinal villi was restored and the structure was clear.3.Compared with the normal control group,the levels of D-LA and DAO in plasma of the model group were significantly increased(P<0.05),and the levels of D-LA decreased after the intervention of aplysin element(P<0.05).The level of DAO decreased significantly(P<0.05).4.Compared with the model group,the concentration of IL-4 in plasma increased(P<0.05).and the level of IL-6 decreased significantly(P<0.05)in the aplysin group.5.The results of Western blotting showed that the expression of occludin and ZO-1 in the model group was significantly lower than that in the normal control group,while the expression of occludin and ZO-1 was significantly increased after the intervention of aplysin.6.The results of RT-PCR showed that there were significant differences in intestinal Escherichia coli,Bacillus faecalis,Clostridium tenella,Lactobacillus and Bifidobacteriu-m between the model group and the normal control group.After the intervention of aplysin,Bacillus faecalis and Bifidobacterium increased in varying degrees,but the difference was not statistically significant.There were significant differences in Escherichia coli,Clostridium tenella and Lactobacillus in the model group.Conclusions1.Aplysin displayed the inhibitory effect on breast cancer rat.2.Aplysin can repair the damaged mucosa,reduce the concentration of D-LA and DAO in plasma,regulate the concentration of inflammatory factors in serum,increase the expression of intestinal TJ protein occludin and ZO-1 and improve the distribution of intestinal flora in rats to fix the intestinal barrier function of the body. |
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